The need to understand the structure of hierarchical or high-dimensional data is present in a variety of fields. Hyperbolic spaces have proven to be an important tool for embedding computations and analysis tasks as their non-linear nature lends itself well to tree or graph data. Subsequently, they have also been used in the visualization of high-dimensional data, where they exhibit increased embedding performance. However, none of the existing dimensionality reduction methods for embedding into hyperbolic spaces scale well with the size of the input data. That is because the embeddings are computed via iterative optimization schemes and the computation cost of every iteration is quadratic in the size of the input. Furthermore, due to the non-linear nature of hyperbolic spaces, euclidean acceleration structures cannot directly be translated to the hyperbolic setting. This article introduces the first acceleration structure for hyperbolic embeddings, building upon a polar quadtree. We compare our approach with existing methods and demonstrate that it computes embeddings of similar quality in significantly less time. Implementation and scripts for the experiments can be found at https://graphics.tudelft.nl/accelerating-hyperbolic-tsne .@en

Exploration and analysis of high-dimensional data are important tasks in many fields that produce large and complex data, like the financial sector, systems biology, or cultural heritage. Tailor-made visual analytics software is developed for each specific application, limiting their applicability in other fields. However, as diverse as these fields are, their characteristics and requirements for data analysis are conceptually similar. Many applications share abstract tasks and data types and are often constructed with similar building blocks. Developing such applications, even when based mostly on existing building blocks, requires significant engineering efforts. We developed ManiVault, a flexible and extensible open-source visual analytics framework for analyzing high-dimensional data. The primary objective of ManiVault is to facilitate rapid prototyping of visual analytics workflows for visualization software developers and practitioners alike. ManiVault is built using a plugin-based architecture that offers easy extensibility. While our architecture deliberately keeps plugins self-contained, to guarantee maximum flexibility and re-usability, we have designed and implemented a messaging API for tight integration and linking of modules to support common visual analytics design patterns. We provide several visualization and analytics plugins, and ManiVault's API makes the integration of new plugins easy for developers. ManiVault facilitates the distribution of visualization and analysis pipelines and results for practitioners through saving and reproducing complete application states. As such, ManiVault can be used as a communication tool among researchers to discuss workflows and results. A copy of this paper and all supplemental material is available at osf.io/9k6jw, and source code at github.com/ManiVaultStudio.

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In spatial transcriptomics (ST) data, biologically relevant features such as tissue compartments or cell-state transitions are reflected by gene expression gradients. Here, we present SpaceWalker, a visual analytics tool for exploring the local gradient structure of 2D and 3D ST data. The user can be guided by the local intrinsic dimensionality of the high-dimensional data to define seed locations, from which a flood-fill algorithm identifies transcriptomically similar cells on the fly, based on the high-dimensional data topology. In several use cases, we demonstrate that the spatial projection of these flooded cells highlights tissue architectural features and that interactive retrieval of gene expression gradients in the spatial and transcriptomic domains confirms known biology. We also show that SpaceWalker generalizes to several different ST protocols and scales well to large, multi-slice, 3D whole-brain ST data while maintaining real-time interaction performance.

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With the rapid advances in single-cell sequencing technologies, novel types of studies into the cell-type makeup of the brain have become possible. Biologists often analyze large and complex single-cell transcriptomic datasets to enhance knowledge of the intricate features of cellular and molecular tissue organization. A particular area of interest is the study of whether cell types and their gene regulation are conserved across species during evolution. However, in-depth comparisons across species of such high-dimensional, multi-modal single-cell data pose considerable visualization challenges. This paper introduces Cytosplore Simian Viewer, a visualization system that combines various views and linked interaction methods for comparative analysis of single-cell transcriptomic datasets across multiple species. Cytosplore Simian Viewer enables biologists to help gain insights into the cell type and gene expression differences and similarities among different species, particularly focusing on comparing human data to other species. The system validation in discovery research on real-world datasets demonstrates its utility in visualizing valuable results related to the evolutionary development of the middle temporal gyrus.@en

The cognitive abilities of humans are distinctive among primates, but their molecular and cellular substrates are poorly understood. We used comparative single-nucleus transcriptomics to analyze samples of the middle temporal gyrus (MTG) from adult humans, chimpanzees, gorillas, rhesus macaques, and common marmosets to understand human-specific features of the neocortex. Human, chimpanzee, and gorilla MTG showed highly similar cell-type composition and laminar organization as well as a large shift in proportions of deep-layer intratelencephalic-projecting neurons compared with macaque and marmoset MTG. Microglia, astrocytes, and oligodendrocytes had more-divergent expression across species compared with neurons or oligodendrocyte precursor cells, and neuronal expression diverged more rapidly on the human lineage. Only a few hundred genes showed human-specific patterning, suggesting that relatively few cellular and molecular changes distinctively define adult human cortical structure.

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High-dimensional images (i.e., with many attributes per pixel) are commonly acquired in many domains, such as geosciences or systems biology. The spatial and attribute information of such data are typically explored separately, e.g., by using coordinated views of an image representation and a low-dimensional embedding of the high-dimensional attribute data. Facing ever growing image data sets, hierarchical dimensionality reduction techniques lend themselves to overcome scalability issues. However, current embedding methods do not provide suitable interactions to reflect image space exploration. Specifically, it is not possible to adjust the level of detail in the embedding hierarchy to reflect changing level of detail in image space stemming from navigation such as zooming and panning. In this paper, we propose such a mapping from image navigation interactions to embedding space adjustments. We show how our mapping applies the "overview first, details-on-demand" characteristic inherent to image exploration in the high-dimensional attribute space. We compare our strategy with regular hierarchical embedding technique interactions and demonstrate the advantages of linking image and embedding interactions through a representative use case.@en

Fiber tracking is a powerful technique that provides valuable insights into the complex white matter structure of the human brain. However, the processing pipeline involves many sources of uncertainty, with one notable factor being the user-defined parameters that significantly influence the resulting outputs. Among these parameters, the definition of seed-points is a crucial aspect in most fiber tracking algorithms. These seed-points are determined through regions of interest (ROI) and serve as the initial points for fiber tract generation. In this work, we present an interactive technique that utilizes seed-point sensitivities to guide the definition of regions of interest (ROI). We examine various scenarios where sensitivity information can enhance the ROI definition process and provide user guidelines and recommended actions for each scenario. Building upon this analysis, we have developed a visualization strategy that enables users to explore seed-point sensitivities effectively and facilitate the definition of optimal ROIs. We present results highlighting the benefits of the proposed visual design in the clinical pipelines.@en

High-dimensional imaging is becoming increasingly relevant in many fields from astronomy and cultural heritage to systems biology. Visual exploration of such high-dimensional data is commonly facilitated by dimensionality reduction. However, common dimensionality reduction methods do not include spatial information present in images, such as local texture features, into the construction of low-dimensional embeddings. Consequently, exploration of such data is typically split into a step focusing on the attribute space followed by a step focusing on spatial information, or vice versa. In this paper, we present a method for incorporating spatial neighborhood information into distance-based dimensionality reduction methods, such as t-Distributed Stochastic Neighbor Embedding (t-SNE). We achieve this by modifying the distance measure between high-dimensional attribute vectors associated with each pixel such that it takes the pixel's spatial neighborhood into account. Based on a classification of different methods for comparing image patches, we explore a number of different approaches. We compare these approaches from a theoretical and experimental point of view. Finally, we illustrate the value of the proposed methods by qualitative and quantitative evaluation on synthetic data and two real-world use cases.

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Microglia have been identified as key players in Alzheimer's disease pathogenesis, and other neurodegenerative diseases. Iba1, and more specifically TMEM119 and P2RY12 are gaining ground as presumedly more specific microglia markers, but comprehensive characterization of the expression of these three markers individually as well as combined is currently missing. Here we used a multispectral immunofluorescence dataset, in which over seventy thousand microglia from both aged controls and Alzheimer patients have been analysed for expression of Iba1, TMEM119 and P2RY12 on a single-cell level. For all markers, we studied the overlap and differences in expression patterns and the effect of proximity to β-amyloid plaques. We found no difference in absolute microglia numbers between control and Alzheimer subjects, but the prevalence of specific combinations of markers (phenotypes) differed greatly. In controls, the majority of microglia expressed all three markers. In Alzheimer patients, a significant loss of TMEM119⁺-phenotypes was observed, independent of the presence of β-amyloid plaques in its proximity. Contrary, phenotypes showing loss of P2RY12, but consistent Iba1 expression were increasingly prevalent around β-amyloid plaques. No morphological features were conclusively associated with loss or gain of any of the markers or any of the identified phenotypes. All in all, none of the three markers were expressed by all microglia, nor can be wholly regarded as a pan- or homeostatic marker, and preferential phenotypes were observed depending on the surrounding pathological or homeostatic environment. This work could help select and interpret microglia markers in previous and future studies.

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Reflectance Imaging Spectroscopy (RIS) is a hyperspectral imaging technique used for investigating the molecular composition of materials. It can help identify pigments used in a painting, which are relevant information for art conservation and history. For every scanned pixel, a reflectance spectrum is obtained and domain experts look for pure representative spectra, called endmembers, which could indicate the presence of particular pigments. However, the identification of endmembers can be a lengthy process, which requires domain experts to manually select pixels and visually inspect multiple spectra in order to find accurate endmembers that belong to the historical context of an investigated painting. We propose an integrated interactive visual-analysis workflow, that combines dimensionality reduction and linked visualizations to identify and inspect endmembers. Here, we present initial results, obtained in collaboration with domain experts.@en

Chronic intestinal inflammation underlies inflammatory bowel disease (IBD). Previous studies indicated alterations in the cellular immune system; however, it has been challenging to interrogate the role of all immune cell subsets simultaneously. Therefore, we aimed to identify immune cell types associated with inflammation in IBD using high-dimensional mass cytometry. We analyzed 188 intestinal biopsies and paired blood samples of newly-diagnosed, treatment-naive patients (n=42) and controls (n=26) in two independent cohorts. We applied mass cytometry (36-antibody panel) to resolve single cells and analyzed the data with unbiased Hierarchical-SNE. In addition, imaging-mass cytometry (IMC) was performed to reveal the spatial distribution of the immune subsets in the tissue. We identified 44 distinct immune subsets. Correlation network analysis identified a network of inflammation-associated subsets, including HLA-DR⁺CD38⁺ EM CD4⁺ T cells, T regulatory-like cells, PD1⁺ EM CD8⁺ T cells, neutrophils, CD27⁺ TCRγδ cells and NK cells. All disease-associated subsets were validated in a second cohort. This network was abundant in a subset of patients, independent of IBD subtype, severity or intestinal location. Putative disease-associated CD4⁺ T cells were detectable in blood. Finally, imaging-mass cytometry revealed the spatial colocalization of neutrophils, memory CD4⁺ T cells and myeloid cells in the inflamed intestine. Our study indicates that a cellular network of both innate and adaptive immune cells colocalizes in inflamed biopsies from a subset of patients. These results contribute to dissecting disease heterogeneity and may guide the development of targeted therapeutics in IBD.

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Controlled human infections provide opportunities to study the interaction between the immune system and malaria parasites, which is essential for vaccine development. Here, we compared immune signatures of malaria-naive Europeans and of Africans with lifelong malaria exposure using mass cytometry, RNA sequencing and data integration, before and 5 and 11 days after venous inoculation with Plasmodium falciparum sporozoites. We observed differences in immune cell populations, antigen-specific responses and gene expression profiles between Europeans and Africans and among Africans with differing degrees of immunity. Before inoculation, an activated/differentiated state of both innate and adaptive cells, including elevated CD161⁺CD4⁺ T cells and interferon-γ production, predicted Africans capable of controlling parasitemia. After inoculation, the rapidity of the transcriptional response and clusters of CD4⁺ T cells, plasmacytoid dendritic cells and innate T cells were among the features distinguishing Africans capable of controlling parasitemia from susceptible individuals. These findings can guide the development of a vaccine effective in malaria-endemic regions.

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Diffusion-Weighted Magnetic Resonance Imaging (DWI) enables the in-vivo visualization of fibrous tissues such as white matter in the brain. Diffusion-Tensor Imaging (DTI) specifically models the DWI diffusion measurements as a second order-tensor. The processing pipeline to visualize this data, from image acquisition to the final rendering, is rather complex. It involves a considerable amount of measurements, parameters and model assumptions, all of which generate uncertainties in the final result which typically are not shown to the analyst in the visualization. In recent years, there has been a considerable amount of work on the visualization of uncertainty in DWI, and specifically DTI. In this chapter, we primarily focus on DTI given its simplicity and applicability, however, several aspects presented are valid for DWI as a whole. We explore the various sources of uncertainties involved, approaches for modeling those uncertainties, and, finally, we survey different strategies to visually represent them. We also look at several related methods of uncertainty visualization that have been applied outside DTI and discuss how these techniques can be adopted to the DTI domain. We conclude our discussion with an overview of potential research directions.@en

Tissue functionality is determined by the characteristics of tissue-resident cells and their interactions within their microenvironment. Imaging Mass Cytometry offers the opportunity to distinguish cell types with high precision and link them to their spatial location in intact tissues at sub-cellular resolution. This technology produces large amounts of spatially-resolved high-dimensional data, which constitutes a serious challenge for the data analysis. We present an interactive visual analysis workflow for the end-to-end analysis of Imaging Mass Cytometry data that was developed in close collaboration with domain expert partners. We implemented the presented workflow in an interactive visual analysis tool; ImaCytE. Our workflow is designed to allow the user to discriminate cell types according to their protein expression profiles and analyze their cellular microenvironments, aiding in the formulation or verification of hypotheses on tissue architecture and function. Finally, we show the effectiveness of our workflow and ImaCytE through a case study performed by a collaborating specialist.@en

Geological analysis of 3D Digital Outcrop Models (DOMs) for reconstruction of ancient habitable environments is a key aspect of the upcoming ESA ExoMars 2022 Rosalind Franklin Rover and the NASA 2020 Rover Perseverance missions in seeking signs of past life on Mars. Geologists measure and interpret 3D DOMs, create sedimentary logs and combine them in ‘correlation panels’ to map the extents of key geological horizons, and build a stratigraphic model to understand their position in the ancient landscape. Currently, the creation of correlation panels is completely manual and therefore time-consuming, and inflexible. With InCorr we present a visualization solution that encompasses a 3D logging tool and an interactive data-driven correlation panel that evolves with the stratigraphic analysis. For the creation of InCorr we closely cooperated with leading planetary geologists in the form of a design study. We verify our results by recreating an existing correlation analysis with InCorr and validate our correlation panel against a manually created illustration. Further, we conducted a user-study with a wider circle of geologists. Our evaluation shows that InCorr efficiently supports the domain experts in tackling their research questions and that it has the potential to significantly impact how geologists work with digital outcrop representations in general.@en

Spatially-resolved omics-data enable researchers to precisely distinguish cell types in tissue and explore their spatial interactions, enabling deep understanding of tissue functionality. To understand what causes or deteriorates a disease and identify related biomarkers, clinical researchers regularly perform large-scale cohort studies, requiring the comparison of such data at cellular level. In such studies, with little a-priori knowledge of what to expect in the data, explorative data analysis is a necessity. Here, we present an interactive visual analysis workflow for the comparison of cohorts of spatially-resolved omics-data. Our workflow allows the comparative analysis of two cohorts based on multiple levels-of-detail, from simple abundance of contained cell types over complex co-localization patterns to individual comparison of complete tissue images. As a result, the workflow enables the identification of cohort-differentiating features, as well as outlier samples at any stage of the workflow. During the development of the workflow, we continuously consulted with domain experts. To show the effectiveness of the workflow, we conducted multiple case studies with domain experts from different application areas and with different data modalities.@en

Diffusion Tensor Imaging (DTI) is a non-invasive magnetic resonance imaging technique that, combined with fiber tracking algorithms, allows the characterization and visualization of white matter structures in the brain. The resulting fiber tracts are used, for example, in tumor surgery to evaluate the potential brain functional damage due to tumor resection. The DTI processing pipeline from image acquisition to the final visualization is rather complex generating undesirable uncertainties in the final results. Most DTI visualization techniques do not provide any information regarding the presence of uncertainty. When planning surgery, a fixed safety margin around the fiber tracts is often used; however, it cannot capture local variability and distribution of the uncertainty, thereby limiting the informed decision-making process. Stochastic techniques are a possibility to estimate uncertainty for the DTI pipeline. However, it has high computational and memory requirements that make it infeasible in a clinical setting. The delay in the visualization of the results adds hindrance to the workflow. We propose a progressive approach that relies on a combination of wild-bootstrapping and fiber tracking to be used within the progressive visual analytics paradigm. We present a local bootstrapping strategy, which reduces the computational and memory costs, and provides fiber-tracking results in a progressive manner. We have also implemented a progressive aggregation technique that computes the distances in the fiber ensemble during progressive bootstrap computations. We present experiments with different scenarios to highlight the benefits of using our progressive visual analytic pipeline in a clinical workflow along with a use case and analysis obtained by discussions with our collaborators.

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Brain iron accumulation has been found to accelerate disease progression in amyloid-β(Aβ) positive Alzheimer patients, though the mechanism is still unknown. Microglia have been identified as key players in the disease pathogenesis, and are highly reactive cells responding to aberrations such as increased iron levels. Therefore, using histological methods, multispectral immunofluorescence and an automated in-house developed microglia segmentation and analysis pipeline, we studied the occurrence of iron-accumulating microglia and the effect on its activation state in human Alzheimer brains. We identified a subset of microglia with increased expression of the iron storage protein ferritin light chain (FTL), together with increased Iba1 expression, decreased TMEM119 and P2RY12 expression. This activated microglia subset represented iron-accumulating microglia and appeared morphologically dystrophic. Multispectral immunofluorescence allowed for spatial analysis of FTL⁺Iba1⁺-microglia, which were found to be the predominant Aβ-plaque infiltrating microglia. Finally, an increase of FTL⁺Iba1⁺-microglia was seen in patients with high Aβ load and Tau load. These findings suggest iron to be taken up by microglia and to influence the functional phenotype of these cells, especially in conjunction with Aβ.

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Imaging mass cytometry (IMC) allows the detection of multiple antigens (approximately 40 markers) combined with spatial information, making it a unique tool for the evaluation of complex biological systems. Due to its widespread availability and retained tissue morphology, formalin-fixed, paraffin-embedded (FFPE) tissues are often a material of choice for IMC studies. However, antibody performance and signal to noise ratios can differ considerably between FFPE tissues as a consequence of variations in tissue processing, including fixation. In contrast to batch effects caused by differences in the immunodetection procedure, variations in tissue processing are difficult to control. We investigated the effect of immunodetection-related signal intensity fluctuations on IMC analysis and phenotype identification, in a cohort of 12 colorectal cancer tissues. Furthermore, we explored different normalization strategies and propose a workflow to normalize IMC data by semi-automated background removal, using publicly available tools. This workflow can be directly applied to previously acquired datasets and considerably improves the quality of IMC data, thereby supporting the analysis and comparison of multiple samples.

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